Glutathione biosynthesis in Arabidopsis trichome cells

In Arabidopsis thaliana, trichome cells are specialized unicellular structures with uncertain functions. Based on earlier observations that one of the genes involved in cysteine biosynthesis (Atcys-3A) is highly expressed in trichomes, we have extended our studies in trichome cells to determine thei...

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Autores: Gutiérrez Alcalá, Gloria, Gotor Martínez, Cecilia, Meyer, Andreas J., Fricker, Mark, Vega, Jorge M., Romero González, Luis Carlos
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2000
País:España
Institución:Universidad de Sevilla (US)
Repositorio:idUS. Depósito de Investigación de la Universidad de Sevilla
OAI Identifier:oai:idus.us.es:11441/69627
Acceso en línea:https://hdl.handle.net/11441/69627
https://doi.org/10.1073/pnas.190334497
Access Level:acceso abierto
Palabra clave:Two-photon laser-scanning microscopy
Sulfate assimilation
O-acetylserine(thiol)lyase
Glutathione
Cysteine
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spelling Glutathione biosynthesis in Arabidopsis trichome cellsGutiérrez Alcalá, GloriaGotor Martínez, CeciliaMeyer, Andreas J.Fricker, MarkVega, Jorge M.Romero González, Luis CarlosTwo-photon laser-scanning microscopySulfate assimilationO-acetylserine(thiol)lyaseGlutathioneCysteineIn Arabidopsis thaliana, trichome cells are specialized unicellular structures with uncertain functions. Based on earlier observations that one of the genes involved in cysteine biosynthesis (Atcys-3A) is highly expressed in trichomes, we have extended our studies in trichome cells to determine their capacity for glutathione (GSH) biosynthesis. First, we have analyzed by in situ hybridization the tissue-specific expression of the genes Atcys-3A and sat5, which encode O-acetylserine(thio)lyase (OASTL) and serine acetyltransferase (SAT), respectively, as well as gsh1 and gsh2, which encode γ-glutamylcysteine synthetase and glutathione synthetase, respectively. The four genes are highly expressed in leaf trichomes of Arabidopsis, and their mRNA accumulate to high levels. Second, we have directly measured cytoplasmic GSH concentration in intact cells by laser-scanning microscopy after labeling with monochlorobimane as a GSH-specific probe. From these measurements, cytosolic GSH concentrations of 238 ± 25, 80 ± 2, and 144 ± 19 μM were estimated for trichome, basement, and epidermal cells, respectively. Taking into account the volume of the cells measured using stereological techniques, the trichomes have a total GSH content more than 300-fold higher than the basement and epidermal cells. Third, after NaCl treatment, GSH biosynthesis is markedly decreased in trichomes. Atcys-3A, sat5, gsh1, and gsh2 mRNA levels show a decrease in transcript abundance, and [GSH](cyt) is reduced to 47 ± 5 μM. These results suggest the important physiological significance of trichome cells related to GSH biosynthesis and their possible role as a sink during detoxification processes.Dirección General de Investigación Científica y Técnica PB96-1367Junta de Andalucía CVI0118National Academy of SciencesQuímica InorgánicaBioquímica Vegetal y Biología MolecularDirección General de Investigación Científica y Técnica (DGICYT). EspañaJunta de Andalucía2000info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttps://hdl.handle.net/11441/69627https://doi.org/10.1073/pnas.190334497reponame:idUS. Depósito de Investigación de la Universidad de Sevillainstname:Universidad de Sevilla (US)InglésProceedings of the National Academy of Sciences of the United States of America, 97 (20), 11108-11113.http://dx.doi.org/10.1073/pnas.190334497info:eu-repo/semantics/openAccessoai:idus.us.es:11441/696272026-06-17T12:51:07Z
dc.title.none.fl_str_mv Glutathione biosynthesis in Arabidopsis trichome cells
title Glutathione biosynthesis in Arabidopsis trichome cells
spellingShingle Glutathione biosynthesis in Arabidopsis trichome cells
Gutiérrez Alcalá, Gloria
Two-photon laser-scanning microscopy
Sulfate assimilation
O-acetylserine(thiol)lyase
Glutathione
Cysteine
title_short Glutathione biosynthesis in Arabidopsis trichome cells
title_full Glutathione biosynthesis in Arabidopsis trichome cells
title_fullStr Glutathione biosynthesis in Arabidopsis trichome cells
title_full_unstemmed Glutathione biosynthesis in Arabidopsis trichome cells
title_sort Glutathione biosynthesis in Arabidopsis trichome cells
dc.creator.none.fl_str_mv Gutiérrez Alcalá, Gloria
Gotor Martínez, Cecilia
Meyer, Andreas J.
Fricker, Mark
Vega, Jorge M.
Romero González, Luis Carlos
author Gutiérrez Alcalá, Gloria
author_facet Gutiérrez Alcalá, Gloria
Gotor Martínez, Cecilia
Meyer, Andreas J.
Fricker, Mark
Vega, Jorge M.
Romero González, Luis Carlos
author_role author
author2 Gotor Martínez, Cecilia
Meyer, Andreas J.
Fricker, Mark
Vega, Jorge M.
Romero González, Luis Carlos
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Química Inorgánica
Bioquímica Vegetal y Biología Molecular
Dirección General de Investigación Científica y Técnica (DGICYT). España
Junta de Andalucía
dc.subject.none.fl_str_mv Two-photon laser-scanning microscopy
Sulfate assimilation
O-acetylserine(thiol)lyase
Glutathione
Cysteine
topic Two-photon laser-scanning microscopy
Sulfate assimilation
O-acetylserine(thiol)lyase
Glutathione
Cysteine
description In Arabidopsis thaliana, trichome cells are specialized unicellular structures with uncertain functions. Based on earlier observations that one of the genes involved in cysteine biosynthesis (Atcys-3A) is highly expressed in trichomes, we have extended our studies in trichome cells to determine their capacity for glutathione (GSH) biosynthesis. First, we have analyzed by in situ hybridization the tissue-specific expression of the genes Atcys-3A and sat5, which encode O-acetylserine(thio)lyase (OASTL) and serine acetyltransferase (SAT), respectively, as well as gsh1 and gsh2, which encode γ-glutamylcysteine synthetase and glutathione synthetase, respectively. The four genes are highly expressed in leaf trichomes of Arabidopsis, and their mRNA accumulate to high levels. Second, we have directly measured cytoplasmic GSH concentration in intact cells by laser-scanning microscopy after labeling with monochlorobimane as a GSH-specific probe. From these measurements, cytosolic GSH concentrations of 238 ± 25, 80 ± 2, and 144 ± 19 μM were estimated for trichome, basement, and epidermal cells, respectively. Taking into account the volume of the cells measured using stereological techniques, the trichomes have a total GSH content more than 300-fold higher than the basement and epidermal cells. Third, after NaCl treatment, GSH biosynthesis is markedly decreased in trichomes. Atcys-3A, sat5, gsh1, and gsh2 mRNA levels show a decrease in transcript abundance, and [GSH](cyt) is reduced to 47 ± 5 μM. These results suggest the important physiological significance of trichome cells related to GSH biosynthesis and their possible role as a sink during detoxification processes.
publishDate 2000
dc.date.none.fl_str_mv 2000
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/11441/69627
https://doi.org/10.1073/pnas.190334497
url https://hdl.handle.net/11441/69627
https://doi.org/10.1073/pnas.190334497
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Proceedings of the National Academy of Sciences of the United States of America, 97 (20), 11108-11113.
http://dx.doi.org/10.1073/pnas.190334497
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv National Academy of Sciences
publisher.none.fl_str_mv National Academy of Sciences
dc.source.none.fl_str_mv reponame:idUS. Depósito de Investigación de la Universidad de Sevilla
instname:Universidad de Sevilla (US)
instname_str Universidad de Sevilla (US)
reponame_str idUS. Depósito de Investigación de la Universidad de Sevilla
collection idUS. Depósito de Investigación de la Universidad de Sevilla
repository.name.fl_str_mv
repository.mail.fl_str_mv
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